Overview | Millipore’sFlowCellect™DualDetectionkitsareaseriesofflowcytometryproductswhichincludeapairofantibodiesthatbindtothesameprotein;onetodetecttotalproteinexpressionandanothertodetectthephosphorylatedformofthesametarget.Usingtwoparameteranalysis,wecanachievetargetspecificdetectionofphosphorylationand,bydoingso,eliminatefalsepositiveswhileenhancingthesignaltonoiseratio.
FlowCellect™HistoneH2A.XDNADamageDualDetectionkitincludestwodirectlyconjugatedantibodies,aphospho-specificAnti-phospho-HistoneH2A.X(Ser139)-PerCPandanAnti-HistoneH2A.X-FITCconjugatedantibodytomeasuretotallevelsofHistoneH2A.X.ThistwocolorkitisdesignedtodetecttheextentofHistoneH2A.XpathwayactivationbymeasuringH2A.XphosphorylationrelativetothetotalH2A.Xexpressioninanygivencellpopulation.Bydoingsuch,thelevelsofboththetotalandphosphorylatedproteincanbemeasuredsimultaneouslyinthesamecell,resultinginanormalizedandaccuratemeasurementofH2A.Xactivationafterstimulation.Moreover,simultaneousmeasurementofbothtotalandphospho-HistoneH2A.Xconfirmstargetspecificityofthephosphorylationevent.Together,atotalandphosphoantibodyduoperformedinmultiplexprovidesanenhancedandmorereliabledetectionofthephospho:totalratiowithinamixedcellpopulation.UsingthisantibodypairprovidesasensitiveandvaluabletooltostudythefactorsthatinduceDNAdamageand/oraffectDNArepair,andallowonetoexplorethelinkagebetweenDNAdamage,cellcyclecheckpoints,andinitiationofapoptosis. |